The anti-proliferative effect of cation channel blockers in T lymphocytes depends on the strength of mitogenic stimulation

• A systematic study of T cell ion channel blockers inhibiting cell proliferation.
• Synergy may exist between ion channel blockers and rapamycin during inhibition.
• Increasing stimulation disrupts the anti-proliferative effect of the blockers.

Ion channels are crucially important for the activation and proliferation of T lymphocytes, and thus, for the function of the immune system. Previous studies on the effects of channel blockers on T cell proliferation reported variable effectiveness due to differing experimental systems. Therefore our aim was to investigate how the strength of the mitogenic stimulation influences the efficiency of cation channel blockers in inhibiting activation, cytokine secretion and proliferation of T cells under standardized conditions.Human peripheral blood lymphocytes were activated via monoclonal antibodies targeting the TCR-CD3 complex and the co-stimulator CD28. We applied the blockers of Kv1.3 (Anuroctoxin), KCa3.1 (TRAM-34) and CRAC (2-Apb) channels of T cells either alone or in combination with rapamycin, the inhibitor of the mammalian target of rapamycin (mTOR). Five days after the stimulation ELISA and flow cytometric measurements were performed to determine IL-10 and IFN-γ secretion, cellular viability and proliferation.Our results showed that ion channel blockers and rapamycin inhibit IL-10 and IFN-γ secretion and cell division in a dose-dependent manner. Simultaneous application of the blockers for each channel along with rapamycin was the most effective, indicating synergy among the various activation pathways. Upon increasing the extent of mitogenic stimulation the anti-proliferative effect of the ion channel blockers diminished. This phenomenon may be important in understanding the fine-tuning of T cell activation.

Ion channel blockers inhibit T lymphocyte proliferation at low mitogen concentrations. This effect diminishes upon using higher mitogen concentrations, but the antiproliferative effect can be recovered by combining ion channel blockers with other immunopharmacological agents such as the mTOR inhibitor rapamycin.Figure optionsDownload as PowerPoint slide